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调节真核基因表达的成分的生物测定：一种参与组蛋白mRNA 3'末端生成的染色体因子。

Bioassay for components regulating eukaryotic gene expression: a chromosomal factor involved in the generation of histone mRNA 3' termini.

作者信息

Stunnenberg H G, Birnstiel M L

出版信息

Proc Natl Acad Sci U S A. 1982 Oct;79(20):6201-4. doi: 10.1073/pnas.79.20.6201.

DOI:10.1073/pnas.79.20.6201
PMID:6959109
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC347087/
Abstract

We have adapted the oocyte injection procedure for the detection of regulatory components involved in the transcription of a eukaryotic mRNA gene. Injection of the histone gene repeat h22 DNA of Psammechinus miliaris into the Xenopus oocyte nucleus results in correct initiation of the histone mRNAs, but readthrough by RNA polymerase occurs at the 3' end of the H3 histone gene (Hentschel, C. C., Probst, E. & Birnstiel, M. L. (1980) Nature (London) 288, 100-102). Coinjection into the oocyte of a chromosomal salt wash fraction derived from sea urchin embryos results in the generation of authentic 3' termini of the histone H3 mRNA. We have partially purified the protein component by column chromatography and density gradient centrifugation. The regulatory factor binds to heparin columns and, hence, has the properties anticipated of an RNA- or DNA-binding protein. The sedimentation coefficient of the active component was determined to be about 12 S, suggesting a molecular weight of 200,000-250,000.

摘要

我们已经调整了卵母细胞注射程序，用于检测参与真核生物mRNA基因转录的调控成分。将沙钱（Psammechinus miliaris）的组蛋白基因重复序列h22 DNA注射到非洲爪蟾卵母细胞核中，可导致组蛋白mRNA正确起始，但RNA聚合酶会在H3组蛋白基因的3'端发生通读（亨切尔，C.C.，普罗布斯特，E.和伯恩施泰尔，M.L.（1980年）《自然》（伦敦）288卷，第100 - 102页）。将源自海胆胚胎的染色体盐洗组分与卵母细胞共同注射，可产生真实的组蛋白H3 mRNA 3'末端。我们通过柱色谱和密度梯度离心对蛋白质成分进行了部分纯化。该调控因子与肝素柱结合，因此具有预期的RNA或DNA结合蛋白的特性。活性成分的沉降系数测定为约12 S，表明分子量为200,000 - 250,000。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/ba47f416cb6c/pnas00459-0097-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/c667ca9c65f4/pnas00459-0096-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/de52383b38e6/pnas00459-0096-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/f9f525fcc960/pnas00459-0097-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/4d299c5e25e5/pnas00459-0097-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/167dfb143d4a/pnas00459-0097-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/ba47f416cb6c/pnas00459-0097-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/c667ca9c65f4/pnas00459-0096-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/de52383b38e6/pnas00459-0096-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/f9f525fcc960/pnas00459-0097-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/4d299c5e25e5/pnas00459-0097-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/167dfb143d4a/pnas00459-0097-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a1ec/347087/ba47f416cb6c/pnas00459-0097-d.jpg

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本文引用的文献

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Delimitation of far upstream sequences required for maximal in vitro transcription of an H2A histone gene.H2A组蛋白基因体外最大转录所需的远上游序列的界定
信使核糖核酸的3'端编辑：序列要求及一段60个核苷酸的核糖核酸在组蛋白信使核糖核酸前体成熟过程中的作用
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